Bukowska Joanna, Ziecik Adam Janusz, Laguna Joanna, Gawronska-Kozak Barbara, Bodek Gabriel
1 In Vitro and Cell Biotechnology Laboratory, Institute of Animal Reproduction and Food Research , Polish Academy of Sciences, Olsztyn, Poland .
2 Department of Hormonal Action Mechanisms, Institute of Animal Reproduction and Food Research , Polish Academy of Sciences, Olsztyn, Poland .
Stem Cells Dev. 2015 Dec 15;24(24):2873-85. doi: 10.1089/scd.2015.0078. Epub 2015 Oct 28.
The regenerative ability of the endometrium is strongly associated with the presence of adult stem/progenitor cells. Purposes of the present study were (1) to establish the presence of stem/progenitor cells in porcine endometrial stroma using a clonogenic assay and (2) to investigate whether the canonical Wnt pathway affects the potential of stem/progenitor cells to undergo self-renewal or differentiation. The utility of endometrial stromal clones as a model for stem/progenitor studies was evaluated based on these cells' increased expression of mesenchymal stem cell (MSC) marker genes, including CD29, CD73, CD90, and CD105, compared with primary cultured cells. Small molecules were introduced to activate (BIO) or inhibit (XAV939) the canonical Wnt pathway during stromal clone formation. Cloning efficiency assays revealed that activation of the Wnt/β-catenin pathway promoted formation of more differentiated small clones. Moreover, activation of the Wnt/β-catenin pathway decreased, whereas inhibition of the pathway increased MSC marker expression. Additionally, we confirmed the importance of canonical Wnt pathway stimulation in endometrial stromal cells through observing the appropriate changes in β-catenin cellular localization. These data indicate that modulation of the canonical Wnt pathway effects the process of regeneration in the porcine endometrium during the course of the estrous cycle.
子宫内膜的再生能力与成体干细胞/祖细胞的存在密切相关。本研究的目的是：（1）使用克隆形成试验确定猪子宫内膜基质中干细胞/祖细胞的存在；（2）研究经典Wnt信号通路是否影响干细胞/祖细胞自我更新或分化的潜能。与原代培养细胞相比，基于这些细胞中包括CD29、CD73、CD90和CD105在内的间充质干细胞（MSC）标记基因表达增加，评估了子宫内膜基质克隆作为干细胞/祖细胞研究模型的实用性。在基质克隆形成过程中引入小分子以激活（BIO）或抑制（XAV939）经典Wnt信号通路。克隆效率分析显示，Wnt/β-连环蛋白信号通路的激活促进了更多分化小克隆的形成。此外，Wnt/β-连环蛋白信号通路的激活降低了，而该信号通路的抑制增加了MSC标记物的表达。另外，我们通过观察β-连环蛋白细胞定位的适当变化，证实了经典Wnt信号通路刺激在子宫内膜基质细胞中的重要性。这些数据表明，经典Wnt信号通路的调节影响了发情周期过程中猪子宫内膜的再生过程。
